Metastic colorectal cancer vaccine

ABSTRACT

The invention relates to prophylactic and therapeutic vaccines for protecting individuals against metastatic colorectal cancer and for treating individuals who are suffering from metastatic colorectal cancer.

FIELD OF THE INVENTION

[0001] The invention relates to prophylactic and therapeutic vaccinesfor protecting individuals against metastatic colorectal cancer and fortreating individuals who are suffering from metastatic colorectalcancer.

BACKGROUND OF THE INVENTION

[0002] Colorectal cancer is the third most common neoplasm worldwide.The mortality rate of newly diagnosed large bowel cancer approaches 50%and there has been little improvement over the past 40 years. Most ofthis mortality reflects local, regional and distant metastases.

[0003] Surgery is the mainstay of treatment for colorectal cancer butrecurrence is frequent. Colorectal cancer has proven resistant tochemotherapy, although limited success has been achieved using acombination of 5-fluorouracil and levamisole. Surgery has had thelargest impact on survival and, in some patients with limited disease,achieves a cure. However, surgery removes bulk tumor, leaving behindmicroscopic residual disease which ultimately results in recrudescence.

[0004] Early detection of primary, metastatic, and recurrent disease cansignificantly impact the prognosis of individuals suffering fromcolorectal cancer. Large bowel cancer diagnosed at an early stage has asignificantly better outcome than that diagnosed at more advancedstages. Similarly, diagnosis of metastatic or recurrent disease earlierpotentially carries with it a better prognosis.

[0005] Recent discoveries have shown that mutations of the human APC(Adenomatous Polyposis Coli) gene are responsible for both sporadic andfamilial colorectal cancers. Germ-line mutations of APC are found ininherited familial cancers such as Gardner's syndrome, attenuatedadenomatous polyposis coli, heredity flat adenoma syndrome and familialadenomatous polyposis (FAP). FAP is an autosomal dominant inheriteddisease predisposing the patient to colon cancer. Patients inheriting asingle mutant allele of APC develop hundreds to thousands of adenomatouspolyps in the second to third decades of life, which if left untreatedprogress to malignant carcinomas. Genetic linkage analysis localized theAPC gene to human chromosome 5q21-q22, a region frequently associatedwith allelic loss of the wildtype 5q allele. Mutations in APC are alsoimplicated in sporadic colorectal cancers and in extracolonic tumors,such as gastric and small intestinal polyps, osteomas, sarcomas anddesmoidal tumors.

[0006] There is a need for improved methods of treating individualssuffering from metastasized colon cancer. There is a need forcompositions useful to treat individuals suffering from metastasizedcolon cancer. There is a need for improved methods of preventing arecurrence of metastasized colon cancer in individuals who have beentreated for metastasized colon cancer. There is a need for compositionsuseful to prevent a recurrence of metastasized colon cancer inindividuals who have been treated for metastasized colon cancer. Thereis a need for improved methods of preventing metastasized colon cancerin individuals, particularly those who have been identified as having agenetic predisposition for colon cancer. There is a need forcompositions useful for preventing metastasized colon cancer inindividuals.

SUMMARY OF THE INVENTION

[0007] The invention relates to an isolated protein comprising at leastone epitope of human ST receptor protein. In some embodiments, theepitope is an epitope of the extracellular domain of the human STreceptor protein. In some embodiments, the epitope is an epitope of thetransmembrane domain of the human ST receptor protein. In someembodiments, the epitope is an epitope of the cytoplasmic domain of thehuman ST receptor protein. In some embodiments, the isolated proteincomprises the extracellular domain of the human ST receptor protein. Insome embodiments, the isolated protein comprises the transmembranedomain of the human ST receptor protein. In some embodiments, theisolated protein comprises the cytoplasmic domain of the human STreceptor protein. In some embodiments, the isolated protein comprisesthe human ST receptor protein. In some embodiments, the isolated proteinconsists of the human ST receptor protein.

[0008] The invention relates to vaccines which comprise such proteinsand a pharmaceutically acceptable carrier or diluent.

[0009] The invention relates to a haptenized protein comprising at leastone epitope of human ST receptor protein. In some embodiments, theepitope is an epitope of the extracellular domain of the human STreceptor protein. In some embodiments, the epitope is an epitope of thetransmembrane domain of the human ST receptor protein. In someembodiments, the epitope is an epitope of the cytoplasmic domain of thehuman ST receptor protein. In some embodiments, the haptenized proteincomprises the extracellular domain of the human ST receptor protein. Insome embodiments, the haptenized protein comprises the transmembranedomain of the human ST receptor protein. In some embodiments, thehaptenized protein comprises the cytoplasmic domain of the human STreceptor protein. In some embodiments, the haptenized protein comprisesthe human ST receptor protein. In some embodiments, the haptenizedprotein consists of the human ST receptor protein.

[0010] The invention relates to vaccines which comprise such haptenizedproteins and a pharmaceutically acceptable carrier or diluent.

[0011] The invention relates to nucleic acid molecules that encode aprotein comprising at least one epitope of human ST receptor protein. Insome embodiments, the nucleic acid molecule encodes a protein with anepitope of the extracellular domain of the human ST receptor protein. Insome embodiments, the nucleic acid molecule encodes a protein with anepitope of the transmembrane domain of the human ST receptor protein. Insome embodiments, the nucleic acid molecule encodes a protein with anepitope of the cytoplasmic domain of the human ST receptor protein. Insome embodiments, the nucleic acid molecule encodes a protein thatprotein comprises the extracellular domain of the human ST receptorprotein. In some embodiments, the nucleic acid molecule encodes aprotein that comprises the transmembrane domain of the human ST receptorprotein. In some embodiments, the nucleic acid molecule encodes aprotein that comprises the cytoplasmic domain of the human ST receptorprotein. In some embodiments, the nucleic acid molecule encodes aprotein that comprises the human ST receptor protein. In someembodiments, the nucleic acid molecule encodes human ST receptorprotein. In some embodiments, the nucleic acid molecule is a plasmid.

[0012] The invention relates to vaccines which comprise such nucleicacid molecules and a pharmaceutically acceptable carrier or diluent.

[0013] The invention relates to vectors that comprise nucleic acidmolecules that encode a protein comprising at least one epitope of humanST receptor protein. In some embodiments, the vector comprises a nucleicacid molecule that encodes a protein with an epitope of theextracellular domain of the human ST receptor protein. In someembodiments, the vector comprises a nucleic acid molecule that encodes aprotein with an epitope of the transmembrane domain of the human STreceptor protein. In some embodiments, the vector comprises a nucleicacid molecule that encodes a protein with an epitope of the cytoplasmicdomain of the human ST receptor protein. In some embodiments, the vectorcomprises a nucleic acid molecule that encodes a protein that proteincomprises the extracellular domain of the human ST receptor protein. Insome embodiments, the vector comprises a nucleic acid molecule thatencodes a protein that comprises the transmembrane domain of the humanST receptor protein. In some embodiments, the vector comprises a nucleicacid molecule that encodes a protein that comprises the cytoplasmicdomain of the human ST receptor protein. In some embodiments, the vectorcomprises a nucleic acid molecule that encodes a protein that comprisesthe human ST receptor protein. In some embodiments, the vector comprisesa nucleic acid molecule that encodes human ST receptor protein. In someembodiments, the vector is a virus or a bacterial cell. In someembodiments, the vector is a recombinant vaccinia virus.

[0014] The invention relates to vaccines which comprise such vectors anda pharmaceutically acceptable carrier or diluent.

[0015] The invention relates to killed or inactivated cells or particlesthat comprise a protein comprising at least one epitope of human STreceptor protein. In some embodiments, the killed or inactivated cellsor particles comprise a protein with an epitope of the extracellulardomain of the human ST receptor protein. In some embodiments, the killedor inactivated cells or particles comprise a protein with an epitope ofthe transmembrane domain of the human ST receptor protein. In someembodiments, the killed or inactivated cells or particles comprise aprotein with an epitope of the cytoplasmic domain of the human STreceptor protein. In some embodiments, the killed or inactivated cellsor particles comprise the extracellular domain of the human ST receptorprotein. In some embodiments, the killed or inactivated cells orparticles vector comprise the transmembrane domain of the human STreceptor protein. In some embodiments, the killed or inactivated cellsor particles comprise the cytoplasmic domain of the human ST receptorprotein. In some embodiments, the killed or inactivated cells orparticles comprise the human ST receptor protein. In some embodiments,the killed or inactivated cells or particles vector is a killed orinactivated colorectal tumor cells.

[0016] The invention relates to vaccines which comprise such killed orinactivated cells or particles and a pharmaceutically acceptable carrieror diluent.

[0017] The invention relates to haptenized killed or inactivated cellsor particles that comprise a protein comprising at least one epitope ofhuman ST receptor protein. In some embodiments, the haptenized killed orinactivated cells or particles comprise a protein with an epitope of theextracellular domain of the human ST receptor protein. In someembodiments, the haptenized killed or inactivated cells or particlescomprise a protein with an epitope of the transmembrane domain of thehuman ST receptor protein. In some embodiments, the haptenized killed orinactivated cells or particles comprise a protein with an epitope of thecytoplasmic domain of the human ST receptor protein. In someembodiments, the haptenized killed or inactivated cells or particlescomprise the extracellular domain of the human ST receptor protein. Insome embodiments, the haptenized killed or inactivated cells orparticles vector comprise the transmembrane domain of the human STreceptor protein. In some embodiments, the haptenized killed orinactivated cells or particles comprise the cytoplasmic domain of thehuman ST receptor protein. In some embodiments, the haptenized killed orinactivated cells or particles comprise the human ST receptor protein.In some embodiments, the haptenized killed or inactivated cells orparticles vector is a killed or inactivated colorectal tumor cells.

[0018] The invention relates to vaccines which comprise such haptenizedkilled or inactivated cells or particles and a pharmaceuticallyacceptable carrier or diluent.

[0019] The present invention relates to methods of treating individualssuffering from metastasized colorectal cancer. The method of the presentinvention provides administering to such an individual a therapeuticallyeffective amount of a vaccine of the invention. The invention relates tothe use of such vaccines as immunotherapeutics.

[0020] The present invention relates to methods of treating individualssusceptible metastasized colorectal cancer. The method of the presentinvention provides administering to such an individual an amount of avaccine of the invention effective to prevent or combat metastasizedcolorectal cancer. The present invention relates to the use of thevaccines of the invention prophylactically.

DETAILED DESCRIPTION OF THE INVENTION

[0021] U.S. Ser. No. 08/141,892 filed on Oct. 26, 1993 (which isscheduled to issue on May 21, 1996 as U.S. Pat. No. 5,518,888), U.S.Ser. No. 08/305,056 filed on Sep. 13, 1994, and PCT Application SerialNumber PCT/US94/12232 filed Oct. 26, 1994, which are each incorporatedherein by reference, describe compositions for and methods of treating,imaging and detecting metastasized colon cancer.

[0022] As used herein, the terms “ST receptor” and “guanylin cyclase C”are interchangeable and meant to refer to the receptors found oncolorectal cells, including local and metastasized colorectal cancercells, which bind to ST. In normal individuals, ST receptors are foundexclusively in cells of intestine, in particular in cells in theduodenum, small intestine (jejunum and ileum), the large intestine,colon (cecum, ascending colon, transverse colon, descending colon andsigmoid colon) and rectum.

[0023] As used herein, the term “colorectal cancer” is meant to includethe well-accepted medical definition that defines colorectal cancer as amedical condition characterized by cancer of cells of the intestinaltract below the small intestine (i.e. the large intestine (colon),including the cecum, ascending colon, transverse colon, descendingcolon, and sigmoid colon, and rectum). Additionally, as used herein, theterm “colorectal cancer” is meant to further include medical conditionswhich are characterized by cancer of cells of the duodenum and smallintestine (jejunum and ileum). The definition of colorectal cancer usedherein is more expansive than the common medical definition but isprovided as such since the cells of the duodenum and small intestinealso contain ST receptors and are therefore amenable to the methods ofthe present invention using the compounds of the present invention.

[0024] As used herein, the term “metastasis” is meant to refer to theprocess in which cancer cells originating in one organ or part of thebody relocate to another part of the body and continue to replicate.Metastasized cells subsequently form tumors which may furthermetastasize. Metastasis thus refers to the spread of cancer from thepart of the body where it originally occurs to other parts of the body.The present invention relates to methods of delivering active agents tometastasized colorectal cancer cells.

[0025] As used herein, the term “metastasized colorectal cancer cells”is meant to refer to colorectal cancer cells which have metastasized;colorectal cancer cells localized in a part of the body other than theduodenum, small intestine (jejunum and ileum), large intestine (colon),including the cecum, ascending colon, transverse colon, descendingcolon, and sigmoid colon, and rectum.

[0026] As used herein, “an individual is suspected of being susceptibleto metastasized colorectal cancer” is meant to refer to an individualwho is at an above-average risk of developing metastasized colorectalcancer. Examples of individuals at a particular risk of developingmetastasized colorectal cancer are those whose family medical historyindicates above average incidence of colorectal cancer among familymembers and/or those who have already developed colorectal cancer andhave been effectively treated who therefore face a risk of relapse andrecurrence. Other factors which may contribute to an above-average riskof developing metastasized colorectal cancer which would thereby lead tothe classification of an individual as being suspected of beingsusceptible to metastasized colorectal cancer may be based upon anindividual's specific genetic, medical and/or behavioral background andcharacteristics.

[0027] Heat stable toxin ST, which is produced by E. coli, as well asother organisms, is responsible for endemic diarrhea in developingcountries and travelers diarrhea. ST induces intestinal secretion bybinding to specific receptors, ST receptors, in the apical brush bordermembranes of the mucosal cells lining the intestinal tract. Binding ofST to ST receptors is non-covalent and occurs in aconcentration-dependent and saturable fashion. Once bound, ST-STreceptor complexes appear to be internalized by intestinal cells, i.e.transported from the surface into the interior of the cell. Binding ofST to ST receptors triggers a cascade of biochemical reactions in theapical membrane of these cells resulting in the production of a signalwhich induces intestinal cells to secrete fluids and electrolytes,resulting in diarrhea.

[0028] ST receptors are unique in that they are only localized in theapical brush border membranes of the cells lining the intestinal tract.Indeed, they are not found in any other cell type in placental mammals.In addition, ST receptors are almost exclusively localized to the apicalmembranes, with little being found in the basolateral membranes on thesides of intestinal cells.

[0029] Mucosal cells lining the intestine are joined together by tightjunctions which form a barrier against the passage of intestinalcontents into the blood stream and components of the blood stream intothe intestinal lumen. Therefore, the apical location of ST receptorsisolates these receptors from the circulatory system so that they may beconsidered to exist separate from the rest of the body; essentially the“outside” of the body. Therefore, the rest of the body is considered“outside” the intestinal tract, i.e. extraintestinal. Compositionsadministered “outside” the intestinal tract are maintained apart andsegregated from the only cells which normally express ST receptors.Conversely, tissue samples taken from tissue outside of the intestinaltract, i.e. extraintestinal tissue samples, do not normally containcells which express ST receptors.

[0030] In individuals suffering from colorectal cancer, the cancer cellsare often derived from cells that produce and display the ST receptorand these cancer cells continue to produce and display the ST receptoron their cell surfaces. Indeed, T84 cells, which are human colonicadenocarcinoma cells isolated from lung metastases, express ST receptorson their cell surface. Similarly, HT29glu-cells, which are human colonicadenocarcinoma cells, express receptors for ST. Thus, in individualssuffering from colorectal cancer, some metastasized intestinal cancercells express ST receptors.

[0031] An effort was undertaken to determine the proportion ofcolorectal tumors which have the ST receptor. Each of the tumors testedwere independently confirmed to be colorectal cancer by standardtechniques of surgical pathology. Every one of the colorectal cancertumors tested, including local colorectal tumors and metastasizedcolorectal tumors (liver, lung, lymph node, peritoneum, ovary) possessedST receptors. In each case, the affinity and density of receptors wasamenable for targeting. Normal liver, lymph node, peritoneum, gallbladder, ovary, stomach, kidney and lung cells were found not to possessST receptors.

[0032] When such cancer cells metastasize, the metastasized cancer cellscontinue to produce and display the ST receptor. The expression of STreceptors on the surfaces of metastatic tumors provides a target whichcan be used to distinguish the metastasized colorectal cancer cells fromnormal extraintestinal cells. This target is useful in the detection,imaging and treatment of metastasized colorectal cancer.

[0033] According to the present invention, the ST receptor proteinserves as a target against which a protective and therapeutic immuneresponse can be induced. Specifically, vaccines are provided whichinduce an immune response against the ST receptor protein. The vaccinesof the invention include, but are not limited to, the following vaccinetechnologies:

[0034] 1) DNA vaccines, i.e. vaccines in which DNA that encodes at leastan epitope from ST receptor protein is administered to an individual'scells where the epitope is expressed and serves as a target for animmune response;

[0035] 2) infectious vector mediated vaccines such as recombinantadenovirus, vaccinia, Salmonella, and BCG wherein the vector carriesgenetic information that encodes at least an epitope of ST receptorprotein such that when the infectious vector is administered to anindividual, the epitope is expressed and serves as a target for animmune response;

[0036] 3) killed or inactivated vaccines which a) comprise either killedcells or inactivated viral particles that display at least an epitope ofthe ST receptor protein and b) when administered to an individual servesas a target for an immune response;

[0037] 3) haptenized killed or inactivated vaccines which a) compriseeither killed cells or inactivated viral particles that display at leastan epitope of the ST receptor, b) are haptenized to be more immunogenicand c) when administered to an individual serves as a target for animmune response;

[0038] 4) subunit vaccines which are vaccines that include proteinmolecules that include at least an epitope the ST receptor protein; and

[0039] 5) haptenized subunit vaccines which are vaccines that a) includeprotein molecules that include at least an epitope the ST receptorprotein and b) are haptenized to be more immunogenic.

[0040] The present invention relates to administering to an individual aprotein or nucleic acid molecule that comprises or encodes,respectively, an immunogenic epitope against which an therapeutic andprophylactic immune response can be induced. Such epitopes are generallyat least 6-8 amino acids in length. The vaccines of the inventiontherefore comprise proteins which are at least, or nucleic acids whichencode at least, 6-8 amino acids in length from ST receptor protein. Thevaccines of the invention may comprise proteins which are at least, ornucleic acids which encode at least, the entire ST receptor protein. Thevaccines of the invention may comprise proteins which are at least, ornucleic acids which encode at least 10 to about 1000 amino acids inlength from ST receptor protein. The vaccines of the invention maycomprise proteins which are at least, or nucleic acids which encode atleast, about 25 to about 500 amino acids in length from ST receptorprotein. The vaccines of the invention may comprise proteins which areat least, or nucleic acids which encode at least, about. 50 to about 400amino acids in length from ST receptor protein. The vaccines of theinvention may comprise proteins which are at least, or nucleic acidswhich encode at least, about 100 to about 300 amino acids in length fromST receptor protein. In preferred embodiments, fragments of ST receptorprotein that include the extracellular domain are provided.

[0041] The present invention relates to compositions for and methods oftreating individuals who are known to have metastasized colorectalcancer. Metastasized colorectal cancer may be diagnosed by those havingordinary skill in the art using art accepted clinical and laboratorypathology protocols and/or those described in U.S. Ser. No. 08/141,892filed on Oct. 26, 1993, U.S. Ser. No. 08/305,056 filed on Sep. 13, 1994,and PCT Application Serial Number PCT/US94/12232 filed Oct. 26, 1994.The present invention provides an immunotherapeutic vaccine useful totreat individuals who have been diagnosed as suffering from metastasizedcolorectal cancer. The immunotherapeutic vaccines of the presentinvention may be administered in combination with other therapiesincluding, but not limited to those described in U.S. Ser. No.08/141,892 filed on Oct. 26, 1993, U.S. Ser. No. 08/305,056 filed onSep. 13, 1994,. and PCT Application Serial Number PCT/US94/12232 filedOct. 26, 1994.

[0042] The present invention relates to compositions for and methods ofpreventing metastatic colorectal cancer in individual is suspected ofbeing susceptible to metastasized colorectal cancer. Such individualsinclude those whose family medical history indicates above averageincidence of colorectal cancer among family members and/or those whohave already developed colorectal cancer and have been effectivelytreated who therefore face a risk of relapse and recurrence. Suchindividuals include those which have been diagnosed as having colorectalcancer including localized only or localized and metastasized colorectalcancer which has been resected or otherwise treated. Such individualsalso include those with an elevated risk as ascertained by geneticevaluation. For example, individuals with APC mutations can beidentified following the U.S. Pat. No. 5,352,775 issued Oct. 4, 1992 toAlbertsen et al., which is incorporated herein by reference.Furthermore, such individuals include: those suffering from inflammatorybowel disease, particularly those with ulcerative colitis; those withcolonic polyps; those with familial adenomatous polyposis, a heritablemutation predisposing patients to develop large numbers of intestinalpolyps; those with Peutz-Jeghers syndrome; those with hereditarynonpolyposis coli, a heritable mutation which predisposes people todevelop colon carcinoma; those with Turcot syndrome-colon carcinoma inconjunction with independent tumors of the central nervous system; andindividuals engaging in rectal intercourse. The vaccines of the presentinvention may be to susceptible individuals prophylactically to preventand combat colorectal cancer metastasis.

[0043] The invention relates to compositions which are the activecomponents of such vaccines or required to make the active components,to methods of making such compositions including the active components,and to methods of making and using vaccines.

[0044] The nucleotide sequence that encodes human ST receptor protein isdisclosed as SEQ ID NO:1. The amino acid sequence of human ST receptoris also disclosed in SEQ ID NO:1. Generally, the extracellular domainrefers to the amino acids about 24 to about 454. The transmembraneregion refers to amino acids about 455 to about 475. The cytoplasmicdomain refers to amino acids about 476 to about 1093.

[0045] Accordingly, some aspects of the invention relate to isolatedproteins that comprise at least one ST receptor epitope. The epitope maybe from the ST receptor extracellular domain, transmembrane domain orcytoplasmic domain. In preferred embodiments, the protein comprises atleast one epitope from the extracellular domain. The protein maycomprise ST receptor protein sequences or consist of ST receptor proteinsequences. The protein may comprise the entire ST receptor protein,consist of the entire ST receptor protein, comprise a fragment of the STreceptor protein, or consist of a fragment of the ST receptor protein.In some preferred embodiments, the protein is a soluble form of theextracellular domain. In some preferred embodiments, the protein is asoluble form of the extracellular domain with a portion of thetransmembrane domain.

[0046] Some aspects of the invention relate to the above describedisolated proteins which are haptenized to render them more immunogenic.That is, some aspects of the invention relate to haptenized proteinsthat comprise at least one ST receptor epitope. The epitope may be fromthe ST receptor extracellular domain, transmembrane domain orcytoplasmic domain. The protein may comprise ST receptor proteinsequences or consist of ST receptor protein sequences. The protein maycomprise the entire ST receptor protein, consist of the entire STreceptor protein, comprise a fragment of the ST receptor protein, orconsist of a fragment of the ST receptor protein. In some preferredembodiments, the haptenized protein comprises a soluble form of theextracellular domain. In some preferred embodiments, the haptenizedprotein is a soluble form of the extracellular domain with a portion ofthe transmembrane domain.

[0047] Some aspects of the invention nucleic acid molecules that encodethe above described isolated proteins.

[0048] Accordingly, some aspects of the invention relate to isolatednucleic acid molecules that encode proteins that comprise at least oneST receptor epitope. The epitope may be from the ST receptorextracellular domain, transmembrane domain or cytoplasmic domain. Inpreferred embodiments, the isolated nucleic acid molecules encodes aprotein that comprises at least one epitope from the extracellulardomain. The isolated nucleic acid molecule may encode a protein thatcomprises or consists of ST receptor protein sequences. The isolatednucleic acid molecule may encode a protein that comprises or consists ofthe entire ST receptor protein, or a protein that comprises or consistsof a fragment of the ST receptor protein. In some embodiments, theisolated nucleic acid molecule encodes non-ST receptor protein sequenceswhich are useful to render the ST receptor protein sequences moreimmunogenic.

[0049] Naked DNA vaccines are described in PCT/US90/01515, which isincorporated herein by reference. Others teach the use of liposomemediated DNA transfer, DNA delivery using microprojectiles (U.S. Pat.No. 4,945,050 issued Jul. 31, 1990 to Sanford et al., which isincorporated herein by reference), and DNA delivery usingelectroporation. In each case, the DNA may be plasmid DNA that isproduced in bacteria, isolated and administered to the animal to betreated. The plasmid DNA molecules are taken up by the cells of theanimal where the sequences that encode the protein of interest areexpressed. The protein thus produced provides a therapeutic orprophylactic effect on the animal.

[0050] The use of vectors including viral vectors and other means ofdelivering nucleic acid molecules to cells of an individual in order toproduce a therapeutic and/or prophylactic immunological effect on theindividual are similarly well known. Recombinant vaccines that employvaccinia vectors are, for example, disclosed in U.S. Pat. No. 5,017,487issued May 21, 1991 to Stunnenberg et al. which is incorporated hereinby reference.

[0051] In some cases, tumor cells from the patient are killed orinactivated and administered as a vaccine product. Berd et al. May 1986Cancer Research 46:2572-2577 and Berd et al. May 1991 Cancer Research51:2731-2734, which are incorporated herein by reference, describes thepreparation and use of tumor cell based vaccine products. According tosome aspects of the present invention, the methods and techniquesdescribed in Berd et al. are adapted by using colorectal cancer cellsinstead of melanoma cells.

[0052] The manufacture and use of subunit vaccines are well known. Onehaving ordinary skill in the art can isolate the nucleic acid moleculethat encode ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof. Once isolated,the nucleic acid molecule can be inserted it into an expression vectorusing standard techniques and readily available starting materials.Rudner et al. May 1995 Proc. Natl. Acad. Sci. USA 92:5169-5173 disclosedthe cloning and expression of the extracellular domain of human STreceptor and purification of the same using a Flag immunoaffinityepitope and antibody therefor.

[0053] The recombinant expression vector that comprises a nucleotidesequence that encodes the nucleic acid molecule that encode ST receptorprotein or a fragment thereof or a protein that comprises the STreceptor protein or a fragment thereof. As used herein, the term“recombinant expression vector” is meant to refer to a plasmid, phage,viral particle or other vector which, when introduced into anappropriate host, contains the necessary genetic elements to directexpression of the coding sequence that encodes the protein. The codingsequence is operably linked to the necessary regulatory sequences.Expression vectors are well known and readily available. Examples ofexpression vectors include plasmids, phages, viral vectors and othernucleic acid molecules or nucleic acid molecule containing vehiclesuseful to transform host cells and facilitate expression of codingsequences. The recombinant expression vectors of the invention areuseful for transforming hosts to prepare recombinant expression systemsfor preparing the isolated proteins of the invention.

[0054] The present invention relates to a host cell that comprises therecombinant expression vector that includes a nucleotide sequence thatencodes the ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof. Host cells foruse in well known recombinant expression systems for production ofproteins are well known and readily available. Examples of host cellsinclude bacteria cells such as E. coli, yeast cells such as S.cerevisiae, insect cells such as S. frugiperda, non-human mammaliantissue culture cells chinese hamster ovary (CHO) cells and human tissueculture cells such as HeLa cells.

[0055] The present invention relates to a transgenic non-human mammalthat comprises the recombinant expression vector that comprises anucleic acid sequence that encodes the proteins of the invention.Transgenic non-human mammals useful to produce recombinant proteins arewell known as are the expression vectors necessary and the techniquesfor generating transgenic animals. Generally, the transgenic animalcomprises a recombinant expression vector in which the nucleotidesequence that encodes the ST receptor protein or a fragment thereof or aprotein that comprises the ST receptor protein or a fragment thereofoperably linked to a mammary cell specific promoter whereby the codingsequence is only expressed in mammary cells and the recombinant proteinso expressed is recovered from the animal's milk.

[0056] In some embodiments, for example, one having ordinary skill inthe art can, using well known techniques, insert such DNA molecules intoa commercially available expression vector for use in well knownexpression systems. For example, the commercially available plasmidpSE420 (Invitrogen, San Diego, Calif.) may be used for production ofcollagen in E. coli. The commercially available plasmid pYES2(Invitrogen, San Diego, Calif.) may, for example, be used for productionin S. cerevisiae strains of yeast. The commercially available MAXBAC™complete baculovirus expression system (Invitrogen, San Diego, Calif.)may, for example, be used for production in insect cells. Thecommercially available plasmid pcDNA I (Invitrogen, San Diego, Calif.)may, for example, be used for production in mammalian cells such asChinese Hamster Ovary cells. One having ordinary skill in the art canuse these commercial expression vectors and systems or others to producethe ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof using routinetechniques and readily available starting materials. (See e.g., Sambrooket al., Molecular Cloning a Laboratory Manual, Second Ed. Cold SpringHarbor Press (1989) which is incorporated herein by reference.) Thus,the desired proteins can be prepared in both prokaryotic and eukaryoticsystems, resulting in a spectrum of processed forms of the protein.

[0057] One having ordinary skill in the art may use other commerciallyavailable expression vectors and systems or produce vectors using wellknown methods and readily available starting materials. Expressionsystems containing the requisite control sequences, such as promotersand polyadenylation signals, and preferably enhancers, are readilyavailable and known in the art for a variety of hosts. See e.g.,Sambrook et al., Molecular Cloning a Laboratory Manual, Second Ed. ColdSpring Harbor Press (1989).

[0058] A wide variety of eukaryotic hosts are also now available forproduction of recombinant foreign proteins. As in bacteria, eukaryotichosts may be transformed with expression systems which produce thedesired protein directly, but more commonly signal sequences areprovided to effect the secretion of the protein. Eukaryotic systems havethe additional advantage that they are able to process introns which mayoccur in the genomic sequences encoding proteins of higher organisms.Eukaryotic systems also provide a variety of processing mechanisms whichresult in, for example, glycosylation, carboxy-terminal amidation,oxidation or derivatization of certain amino acid residues,conformational control, and so forth.

[0059] Commonly used eukaryotic systems include, but is not limited to,yeast, fungal cells, insect cells, mammalian cells, avian cells, andcells of higher plants. Suitable promoters are available which arecompatible and operable for use in each of these host types as well asare termination sequences and enhancers, e.g. the baculovirus polyhedronpromoter. As above, promoters can be either constitutive or inducible.For example, in mammalian systems, the mouse metallothionein promotercan be induced by the addition of heavy metal ions.

[0060] The particulars for the construction of expression systemssuitable for desired hosts are known to those in the art. Briefly, forrecombinant production of the protein, the DNA encoding the polypeptideis suitably ligated into the expression vector of choice. The DNA isoperably linked to all regulatory elements which are necessary forexpression of the DNA in the selected host. One having ordinary skill inthe art can, using well known techniques, prepare expression vectors forrecombinant production of the polypeptide.

[0061] The expression vector including the DNA that encodes the STreceptor protein or a fragment thereof or a protein that comprises theST receptor protein or a fragment thereof is used to transform thecompatible host which is then cultured and maintained under conditionswherein expression of the foreign DNA takes place. The protein of thepresent invention thus produced is recovered from the culture, either bylysing the cells or from the culture medium as appropriate and known tothose in the art. One having ordinary skill in the art can, using wellknown techniques, the ST receptor protein or a fragment thereof or aprotein that comprises the ST receptor protein or a fragment thereofthat is produced using such expression systems. The methods of purifyingthe ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof using antibodieswhich specifically bind to the protein are well known. Antibodies whichspecifically bind to a particular protein may be used to purify theprotein from natural sources using well known techniques and readilyavailable starting materials. Such antibodies may also be used to purifythe protein from material present when producing the protein byrecombinant DNA methodology. The present invention relates to antibodiesthat bind to an epitope which is present on the ST receptor protein or afragment thereof or a protein that comprises the ST receptor protein ora fragment thereof. As used herein, the term “antibody” is meant torefer to complete, intact antibodies, and Fab fragments and F(ab)₂fragments thereof. Complete, intact antibodies include monoclonalantibodies such as murine monoclonal antibodies, chimeric antibodies andhumanized antibodies. Antibodies that bind to an epitope which ispresent on the ST receptor protein or a fragment thereof or a proteinthat comprises the ST receptor protein or a fragment thereof are usefulto isolate and purify the protein from both natural sources orrecombinant expression systems using well known techniques such asaffinity chromatography. Immunoaffinity techniques generally aredescribed in Waldman et al. 1991 Methods of Enzymol. 195:391-396, whichis incorporated herein by reference. Antibodies are useful to detect thepresence of such protein in a sample and to determine if cells areexpressing the protein. The production of antibodies and the proteinstructures of complete, intact antibodies, Fab fragments and F(ab)₂fragments and the organization of the genetic sequences that encode suchmolecules are well known and are described, for example, in Harlow, E.and D. Lane (1988) ANTIBODIES: A Laboratory Manual, Cold Spring HarborLaboratory, Cold Spring Harbor, N.Y. which is incorporated herein byreference. Briefly, for example, the ST receptor protein or a fragmentthereof or a protein that comprises the ST receptor protein or afragment thereof, or an immunogenic fragment thereof is injected intomice. The spleen of the mouse is removed, the spleen cells are isolatedand fused with immortalized mouse cells. The hybrid cells, orhybridomas, are cultured and those cells which secrete antibodies areselected. The antibodies are analyzed and, if found to specifically bindto the ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof, the hybridomawhich produces them is cultured to produce a continuous supply ofantibodies.

[0062] In some embodiments of the invention, transgenic non-humananimals are generated. The transgenic animals according to the inventioncontain nculeotides that encode the ST receptor protein or a fragmentthereof or a protein that comprises the ST receptor protein or afragment thereof under the regulatory control of a mammary specificpromoter. One having ordinary skill in the art using standardtechniques, such as those taught in U.S. Pat. No. 4,873,191 issued Oct.10, 1989 to Wagner and U.S. Pat. No. 4,736,866 issued Apr. 12, 1988 toLeder, both of which are incorporated herein by reference, can producetransgenic animals which produce the ST receptor protein or a fragmentthereof or a protein that comprises the ST receptor protein or afragment thereof. Preferred animals are goats and rodents, particularlyrats and mice.

[0063] In addition to producing these proteins by recombinanttechniques, automated peptide synthesizers may also be employed toproduce the ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof of theinvention. Such techniques are well known to those having ordinary skillin the art and are useful if derivatives which have substitutions notprovided for in DNA-encoded protein production.

[0064] In some embodiments, the protein that makes up a subunit vaccineor the cells or particles of a killed or inactivated vaccine may behaptenized to increase immunogenicity. In some cases, the haptenizationis the conjugation of a larger molecular structure to the ST receptorprotein or a fragment thereof or a protein that comprises the STreceptor protein or a fragment thereof. In some cases, tumor cells fromthe patient are killed and haptenized as a means to make an effectivevaccine product. In cases in which other cells, such as bacteria oreukaryotic cells which are provided with the genetic information to makeand display the ST receptor protein or a fragment thereof or a proteinthat comprises the ST receptor protein or a fragment thereof, are killedand used as the active vaccine component, such cells are haptenized toincrease immunogenicity. Haptenization is well known and can be readilyperformed.

[0065] Methods of haptenizing cells generally and tumor cells inparticular are described in Berd et al. May 1986 Cancer Research46:2572-2577 and Berd et al. May 1991 Cancer Research 51:2731-2734,which are incorporated herein by reference. Additional haptenizationprotocols are disclosed in Miller et al. 1976 J. Immunol.117(5:1):1591-1526.

[0066] Haptenization compositions and methods which may be adapted to beused to prepare haptenized ST immunogens according to the presentinvention include those described in the following U.S. Patents whichare each incorporated herein by reference: U.S. Pat. No. 5,037,645issued Aug. 6, 1991 to Strahilevitz; U.S. Pat. No. 5,112,606 issued May12, 1992 to Shiosaka et al.; U.S. Pat. No. 4,526716 issued Jul. 2, 1985to Stevens; U.S. Pat. No. 4,329,281 issued May 11, 1982 to Christensonet al.; and U.S. Pat. No. 4,022,878 issued May 10, 1977 to Gross.Peptide vaccines and methods of enhancing immunogenicity of peptideswhich may be adapted to modify ST immunogens of the invention are alsodescribed in Francis et al. 1989 Methods of Enzymol. 178:659-676, whichis incorporated herein by reference. Sad et al. 1992 Immunolology76:599-603, which is incorporated herein by reference, teaches methodsof making immunotherapeutic vaccines by conjugating gonadotropinreleasing hormone to diphtheria toxoid. ST immunogens may be similarlyconjugated to produce an immunotherapeutic vaccine of the presentinvention. MacLean et al. 1993 Cancer Immunol. Immunother. 36:215-22.2,which is incorporated herein by reference, describes conjugationmethodologies for producing immunotherapeutic vaccines which may beadaptable to produce an immunotherapeutic vaccine of the presentinvention. The hapten is keyhole limpet hemocyanin which may beconjugated to an ST immunogen.

[0067] As used herein, the term “ST receptor immunogen” is meant torefer to the ST receptor protein or a fragment thereof or a protein thatcomprises the ST receptor protein or a fragment thereof, haptenized STreceptor protein or a haptenized fragment thereof or a haptenizedprotein that comprises the ST receptor protein or a haptenized fragmentthereof, cells and particles which display at least one ST receptorepitope, and haptenized cells and haptenized particles which display atleast one ST receptor epitope

[0068] Vaccines according to some aspects of the invention comprise apharmaceutically acceptable carrier in combination with an ST receptorimmunogen. Pharmaceutical formulations are well known and pharmaceuticalcompositions comprising such proteins may be routinely formulated by onehaving ordinary skill in the art. Suitable pharmaceutical carriers aredescribed in Remington's Pharmaceutical Sciences, A. Osol, a standardreference text in this field, which is incorporated herein by reference.The present invention relates to an injectable pharmaceuticalcomposition that comprises a pharmaceutically acceptable carrier and anST receptor immunogen. The ST receptor immunogen is preferably sterileand combined with a sterile pharmaceutical carrier.

[0069] In some embodiments, for example, the ST receptor immunogen canbe formulated as a solution, suspension, emulsion or lyophilized powderin association with a pharmaceutically acceptable vehicle. Examples ofsuch vehicles are water, saline, Ringer's solution, dextrose solution,and 5% human serum albumin. Liposomes and nonaqueous vehicles such asfixed oils may also be used. The vehicle or lyophilized powder maycontain additives that maintain isotonicity (e.g., sodium chloride,mannitol) and chemical stability (e.g., buffers and preservatives). Theformulation is sterilized by commonly used techniques.

[0070] An injectable composition may comprise the ST receptor immunogenin a diluting agent such as, for example, sterile water,electrolytes/dextrose, fatty oils of vegetable origin, fatty esters, orpolyols, such as propylene glycol and polyethylene glycol. Theinjectable must be sterile and free of pyrogens.

[0071] The vaccines of the present invention may be administered by anymeans that enables the immunogenic agent to be presented to the body'simmune system for recognition and induction of an immunogenic response.Pharmaceutical compositions may be administered parenterally, i.e.,intravenous, subcutaneous, intramuscular.

[0072] Dosage varies depending upon known factors such as thepharmacodynamic characteristics of the particular agent, and its modeand route of administration; age, health, and weight of the recipient;nature and extent of symptoms, kind of concurrent treatment, frequencyof treatment, and the effect desired. An amount of immunogen isdelivered to induce a protective or therapeutically effective immuneresponse. Those having ordinary skill in the art can readily determinethe range and optimal dosage by routine methods.

1 2 1 3786 DNA Homo sapiens CDS (118)..(3336) 1 tggagtgggc tgagggactccactagaggc tgtccatctg gattccctgc ctccctagga 60 gcccaacaga gcaaagcaagtgggcacaag gagtatggtt ctaacgtgat tggggtc 117 y atg aag acg ttg ctg ttggac ttg gct ttg tgg tca ctg ctc ttc cag 165 Met Lys Thr Leu Leu Leu AspLeu Ala Leu Trp Ser Leu Leu Phe Gln 1 5 10 15 y ccc ggg tgg ctg tcc tttagt tcc cag gtg agt cag aac tgc cac aat 213 Pro Gly Trp Leu Ser Phe SerSer Gln Val Ser Gln Asn Cys His Asn 20 25 30 y ggc agc tat gaa atc agcgtc ctg atg atg ggc aac tca gcc ttt gca 261 Gly Ser Tyr Glu Ile Ser ValLeu Met Met Gly Asn Ser Ala Phe Ala 35 40 45 y gag ccc ctg aaa aac ttggaa gat gcg gtg aat gag ggg ctg gaa ata 309 Glu Pro Leu Lys Asn Leu GluAsp Ala Val Asn Glu Gly Leu Glu Ile 50 55 60 y gtg aga gga cgt ctg caaaat gct ggc cta aat gtg act gtg aac gct 357 Val Arg Gly Arg Leu Gln AsnAla Gly Leu Asn Val Thr Val Asn Ala 65 70 75 80 y act ttc atg tat tcggat ggt ctg att cat aac tca ggc gac tgc cgg 405 Thr Phe Met Tyr Ser AspGly Leu Ile His Asn Ser Gly Asp Cys Arg 85 90 95 y agt agc acc tgt gaaggc ctc gac cta ctc agg aaa att tca aat gca 453 Ser Ser Thr Cys Glu GlyLeu Asp Leu Leu Arg Lys Ile Ser Asn Ala 100 105 110 y caa cgg atg ggctgt gtc ctc ata ggg ccc tca tgt aca tac tcc acc 501 Gln Arg Met Gly CysVal Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr 115 120 125 y ttc cag atgtac ctt gac aca gaa ttg agc tac ccc atg atc tca gct 549 Phe Gln Met TyrLeu Asp Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala 130 135 140 y gga agtttt gga ttg tca tgt gac tat aaa gaa acc tta acc agg ctg 597 Gly Ser PheGly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu 145 150 155 160 yatg tct cca gct aga aag ttg atg tac ttc ttg gtt aac ttt tgg aaa 645 MetSer Pro Ala Arg Lys Leu Met Tyr Phe Leu Val Asn Phe Trp Lys 165 170 175y acc aac gat ctg ccc ttc aaa act tat tcc tgg agc act tcg tat gtt 693Thr Asn Asp Leu Pro Phe Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val 180 185190 y tac aag aat ggt aca gaa act gag gac tgt ttc tgg tac ctt aat gct741 Tyr Lys Asn Gly Thr Glu Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala 195200 205 y ctg gag gct agc gtt tcc tat ttc tcc cac gaa ctc ggc ttt aaggtg 789 Leu Glu Ala Ser Val Ser Tyr Phe Ser His Glu Leu Gly Phe Lys Val210 215 220 y gtg tta aga caa gat aag gag ttt cag gat atc tta atg gaccac aac 837 Val Leu Arg Gln Asp Lys Glu Phe Gln Asp Ile Leu Met Asp HisAsn 225 230 235 240 y agg aaa agc aat gtg att att atg tgt ggt ggt ccagag ttc ctc tac 885 Arg Lys Ser Asn Val Ile Ile Met Cys Gly Gly Pro GluPhe Leu Tyr 245 250 255 y aag ctg aag ggt gac cga gca gtg gct gaa gacatt gtc att att cta 933 Lys Leu Lys Gly Asp Arg Ala Val Ala Glu Asp IleVal Ile Ile Leu 260 265 270 y gtg gat ctt ttc aat gac cag tac ttg gaggac aat gtc aca gcc cct 981 Val Asp Leu Phe Asn Asp Gln Tyr Leu Glu AspAsn Val Thr Ala Pro 275 280 285 gac tat atg aaa aat gtc ctt gtt ctg acgctg tct cct ggg aat tcc 1029 Asp Tyr Met Lys Asn Val Leu Val Leu Thr LeuSer Pro Gly Asn Ser 290 295 300 ctt cta aat agc tct ttc tcc agg aat ctatca cca aca aaa cga gac 1077 Leu Leu Asn Ser Ser Phe Ser Arg Asn Leu SerPro Thr Lys Arg Asp 305 310 315 320 ttt cgt ctt gcc tat ttg aat gga atcctc gtc ttt gga cat atg ctg 1125 Phe Arg Leu Ala Tyr Leu Asn Gly Ile LeuVal Phe Gly His Met Leu 325 330 335 aag ata ttt ctt gaa aat gga gaa aatatt acc acc ccc aaa ttt gct 1173 Lys Ile Phe Leu Glu Asn Gly Glu Asn IleThr Thr Pro Lys Phe Ala 340 345 350 cat gcc ttc agg aat ctc act ttt gaaggg tat gac ggt cca gtg acc 1221 His Ala Phe Arg Asn Leu Thr Phe Glu GlyTyr Asp Gly Pro Val Thr 355 360 365 ttg gat gac tgg ggg gat gtt gac agtacc atg gtg ctt ctg tat acc 1269 Leu Asp Asp Trp Gly Asp Val Asp Ser ThrMet Val Leu Leu Tyr Thr 370 375 380 tct gtg gac acc aag aaa tac aag gttctt ttg acc tat gat acc cac 1317 Ser Val Asp Thr Lys Lys Tyr Lys Val LeuLeu Thr Tyr Asp Thr His 385 390 395 400 gta aat aag acc tat cct gtg gatatg agc ccc aca ttc act tgg aag 1365 Val Asn Lys Thr Tyr Pro Val Asp MetSer Pro Thr Phe Thr Trp Lys 405 410 415 aac tct aaa ctt cct aat gat attaca ggc cgg ggc cct cag atc ctg 1413 Asn Ser Lys Leu Pro Asn Asp Ile ThrGly Arg Gly Pro Gln Ile Leu 420 425 430 atg att gca gtc ttc acc ctc actgga gct gtg gtg ctg ctc ctg ctc 1461 Met Ile Ala Val Phe Thr Leu Thr GlyAla Val Val Leu Leu Leu Leu 435 440 445 gtc gct ctc ctg atg ctc aga aaatat aga aaa gat tat gaa ctt cgt 1509 Val Ala Leu Leu Met Leu Arg Lys TyrArg Lys Asp Tyr Glu Leu Arg 450 455 460 cag aaa aaa tgg tcc cac att cctcct gaa aat atc ttt cct ctg gag 1557 Gln Lys Lys Trp Ser His Ile Pro ProGlu Asn Ile Phe Pro Leu Glu 465 470 475 480 acc aat gag acc aat cat gttagc ctc aag atc gat gat gac aaa aga 1605 Thr Asn Glu Thr Asn His Val SerLeu Lys Ile Asp Asp Asp Lys Arg 485 490 495 cga gat aca atc cag aga ctacga cag tgc aaa tac gtc aaa aag cga 1653 Arg Asp Thr Ile Gln Arg Leu ArgGln Cys Lys Tyr Val Lys Lys Arg 500 505 510 gtg att ctc aaa gat ctc aagcac aat gat ggt aat ttc act gaa aaa 1701 Val Ile Leu Lys Asp Leu Lys HisAsn Asp Gly Asn Phe Thr Glu Lys 515 520 525 cag aag ata gaa ttg aac aagttg ctt cag att gac tat tac acc cta 1749 Gln Lys Ile Glu Leu Asn Lys LeuLeu Gln Ile Asp Tyr Tyr Thr Leu 530 535 540 acc aag ttc tac ggg aca gtgaaa ctg gat acc atg atc ttc ggg gtg 1797 Thr Lys Phe Tyr Gly Thr Val LysLeu Asp Thr Met Ile Phe Gly Val 545 550 555 560 ata gaa tac tgt gag agagga tcc ctc cgg gaa gtt tta aat gac aca 1845 Ile Glu Tyr Cys Glu Arg GlySer Leu Arg Glu Val Leu Asn Asp Thr 565 570 575 att tcc tac cct gat ggcaca ttc atg gat tgg gag ttt aag atc tct 1893 Ile Ser Tyr Pro Asp Gly ThrPhe Met Asp Trp Glu Phe Lys Ile Ser 580 585 590 gtc ttg tat gac att gctaag gga atg tca tat ctg cac tcc agt aag 1941 Val Leu Tyr Asp Ile Ala LysGly Met Ser Tyr Leu His Ser Ser Lys 595 600 605 aca gaa gtc cat ggt cgtctg aaa tct acc aac tgc gta gtg gac agt 1989 Thr Glu Val His Gly Arg LeuLys Ser Thr Asn Cys Val Val Asp Ser 610 615 620 aga atg gtg gtg aag atcact gat ttt ggc tgc aat tcc att ttg cct 2037 Arg Met Val Val Lys Ile ThrAsp Phe Gly Cys Asn Ser Ile Leu Pro 625 630 635 640 cca aaa aag gac ctgtgg aca gct cca gag cac ctc cgc caa gcc aac 2085 Pro Lys Lys Asp Leu TrpThr Ala Pro Glu His Leu Arg Gln Ala Asn 645 650 655 atc tct cag aaa ggagat gtg tac agc tat ggg atc atc gca cag gag 2133 Ile Ser Gln Lys Gly AspVal Tyr Ser Tyr Gly Ile Ile Ala Gln Glu 660 665 670 atc att ctg cgg aaagaa acc ttc tac act ttg agc tgt cgg gac cgg 2181 Ile Ile Leu Arg Lys GluThr Phe Tyr Thr Leu Ser Cys Arg Asp Arg 675 680 685 aat gag aag att ttcaga gtg gaa aat tcc aat gga atg aaa ccc ttc 2229 Asn Glu Lys Ile Phe ArgVal Glu Asn Ser Asn Gly Met Lys Pro Phe 690 695 700 cgc cca gat tta ttcttg gaa aca gca gag gaa aaa gag cta gaa gtg 2277 Arg Pro Asp Leu Phe LeuGlu Thr Ala Glu Glu Lys Glu Leu Glu Val 705 710 715 720 tac cta ctt gtaaaa aac tgt tgg gag gaa gat cca gaa aag aga cca 2325 Tyr Leu Leu Val LysAsn Cys Trp Glu Glu Asp Pro Glu Lys Arg Pro 725 730 735 gat ttc aaa aaaatt gag act aca ctt gcc aag ata ttt gga ctt ttt 2373 Asp Phe Lys Lys IleGlu Thr Thr Leu Ala Lys Ile Phe Gly Leu Phe 740 745 750 cat gac caa aaaaat gaa agc tat atg gat acc ttg atc cga cgt cta 2421 His Asp Gln Lys AsnGlu Ser Tyr Met Asp Thr Leu Ile Arg Arg Leu 755 760 765 cag cta tat tctcga aac ctg gaa cat ctg gta gag gaa agg aca cag 2469 Gln Leu Tyr Ser ArgAsn Leu Glu His Leu Val Glu Glu Arg Thr Gln 770 775 780 ctg tac aag gcagag agg gac agg gct gac aga ctt aac ttt atg ttg 2517 Leu Tyr Lys Ala GluArg Asp Arg Ala Asp Arg Leu Asn Phe Met Leu 785 790 795 800 ctt cca aggcta gtg gta aag tct ctg aag gag aaa ggc ttt gtg gag 2565 Leu Pro Arg LeuVal Val Lys Ser Leu Lys Glu Lys Gly Phe Val Glu 805 810 815 ccg gaa ctatat gag gaa gtt aca atc tac ttc agt gac att gta ggt 2613 Pro Glu Leu TyrGlu Glu Val Thr Ile Tyr Phe Ser Asp Ile Val Gly 820 825 830 ttc act actatc tgc aaa tac agc acc ccc atg gaa gtg gtg gac atg 2661 Phe Thr Thr IleCys Lys Tyr Ser Thr Pro Met Glu Val Val Asp Met 835 840 845 ctt aat gacatc tat aag agt ttt gac cac att gtt gat cat cat gat 2709 Leu Asn Asp IleTyr Lys Ser Phe Asp His Ile Val Asp His His Asp 850 855 860 gtc tac aaggtg gaa acc atc ggt gat gcg tac atg gtg gct agt ggt 2757 Val Tyr Lys ValGlu Thr Ile Gly Asp Ala Tyr Met Val Ala Ser Gly 865 870 875 880 ttg cctaag aga aat ggc aat cgg cat gca ata gac att gcc aag atg 2805 Leu Pro LysArg Asn Gly Asn Arg His Ala Ile Asp Ile Ala Lys Met 885 890 895 gcc ttggaa atc ctc agc ttc atg ggg acc ttt gag ctg gag cat ctt 2853 Ala Leu GluIle Leu Ser Phe Met Gly Thr Phe Glu Leu Glu His Leu 900 905 910 cct ggcctc cca ata tgg att cgc att gga gtt cac tct ggt ccc tgt 2901 Pro Gly LeuPro Ile Trp Ile Arg Ile Gly Val His Ser Gly Pro Cys 915 920 925 gct gctgga gtt gtg gga atc aag atg cct cgt tat tgt cta ttt gga 2949 Ala Ala GlyVal Val Gly Ile Lys Met Pro Arg Tyr Cys Leu Phe Gly 930 935 940 gat acggtc aac aca gcc tct agg atg gaa tcc act ggc ctc cct ttg 2997 Asp Thr ValAsn Thr Ala Ser Arg Met Glu Ser Thr Gly Leu Pro Leu 945 950 955 960 agaatt cac gtg agt ggc tcc acc ata gcc atc ctg aag aga act gag 3045 Arg IleHis Val Ser Gly Ser Thr Ile Ala Ile Leu Lys Arg Thr Glu 965 970 975 tgccag ttc ctt tat gaa gtg aga gga gaa aca tac tta aag gga aga 3093 Cys GlnPhe Leu Tyr Glu Val Arg Gly Glu Thr Tyr Leu Lys Gly Arg 980 985 990 ggaaat gag act acc tac tgg ctg act ggg atg aag gac cag aaa ttc 3141 Gly AsnGlu Thr Thr Tyr Trp Leu Thr Gly Met Lys Asp Gln Lys Phe 995 1000 1005aac ctg cca acc cct cct act gtg gag aat caa cag cgt ttg caa gca 3189 AsnLeu Pro Thr Pro Pro Thr Val Glu Asn Gln Gln Arg Leu Gln Ala 1010 10151020 gaa ttt tca gac atg att gcc aac tct tta cag aaa aga cag gca gca3237 Glu Phe Ser Asp Met Ile Ala Asn Ser Leu Gln Lys Arg Gln Ala Ala1025 1030 1035 1040 ggg ata aga agc caa aaa ccc aga cgg gta gcc agc tataaa aaa ggc 3285 Gly Ile Arg Ser Gln Lys Pro Arg Arg Val Ala Ser Tyr LysLys Gly 1045 1050 1055 act ctg gaa tac ttg cag ctg aat acc aca gac aaggag agc acc tat 3333 Thr Leu Glu Tyr Leu Gln Leu Asn Thr Thr Asp Lys GluSer Thr Tyr 1060 1065 1070 ttt taaacctaaa tgaggtataa ggactcacacaaattaaaat acagctgcac 3386 Phe tgaggccagg caccctcagg tgtcctgaaagcttactttc ctgagacctc atgaggcaga 3446 aatgtcttag gcttggctgc cctgtttggaccatggactt tctttgcatg aatcagatgt 3506 gttctcagtg aaataactac cttccactctggaaccttat tccagcagtt gttccaggga 3566 gcttctacct ggaaaagaaa agaatttcatttattttttg tttgtttatt tttatcgttt 3626 ttgtttactg gctttccttc tgtattcataagatttttta aattgtcata attatatttt 3686 aaatacccat cttcattaaa gtatatttaactcataattt ttgcagaaaa tatgctatat 3746 attaggcaag aataaaagct aaaggtttcccaaaaaaaaa 3786 2 1073 PRT Homo sapiens 2 Met Lys Thr Leu Leu Leu AspLeu Ala Leu Trp Ser Leu Leu Phe Gln 1 5 10 15 Pro Gly Trp Leu Ser PheSer Ser Gln Val Ser Gln Asn Cys His Asn 20 25 30 Gly Ser Tyr Glu Ile SerVal Leu Met Met Gly Asn Ser Ala Phe Ala 35 40 45 Glu Pro Leu Lys Asn LeuGlu Asp Ala Val Asn Glu Gly Leu Glu Ile 50 55 60 Val Arg Gly Arg Leu GlnAsn Ala Gly Leu Asn Val Thr Val Asn Ala 65 70 75 80 Thr Phe Met Tyr SerAsp Gly Leu Ile His Asn Ser Gly Asp Cys Arg 85 90 95 Ser Ser Thr Cys GluGly Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala 100 105 110 Gln Arg Met GlyCys Val Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr 115 120 125 Phe Gln MetTyr Leu Asp Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala 130 135 140 Gly SerPhe Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu 145 150 155 160Met Ser Pro Ala Arg Lys Leu Met Tyr Phe Leu Val Asn Phe Trp Lys 165 170175 Thr Asn Asp Leu Pro Phe Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val 180185 190 Tyr Lys Asn Gly Thr Glu Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala195 200 205 Leu Glu Ala Ser Val Ser Tyr Phe Ser His Glu Leu Gly Phe LysVal 210 215 220 Val Leu Arg Gln Asp Lys Glu Phe Gln Asp Ile Leu Met AspHis Asn 225 230 235 240 Arg Lys Ser Asn Val Ile Ile Met Cys Gly Gly ProGlu Phe Leu Tyr 245 250 255 Lys Leu Lys Gly Asp Arg Ala Val Ala Glu AspIle Val Ile Ile Leu 260 265 270 Val Asp Leu Phe Asn Asp Gln Tyr Leu GluAsp Asn Val Thr Ala Pro 275 280 285 Asp Tyr Met Lys Asn Val Leu Val LeuThr Leu Ser Pro Gly Asn Ser 290 295 300 Leu Leu Asn Ser Ser Phe Ser ArgAsn Leu Ser Pro Thr Lys Arg Asp 305 310 315 320 Phe Arg Leu Ala Tyr LeuAsn Gly Ile Leu Val Phe Gly His Met Leu 325 330 335 Lys Ile Phe Leu GluAsn Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala 340 345 350 His Ala Phe ArgAsn Leu Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr 355 360 365 Leu Asp AspTrp Gly Asp Val Asp Ser Thr Met Val Leu Leu Tyr Thr 370 375 380 Ser ValAsp Thr Lys Lys Tyr Lys Val Leu Leu Thr Tyr Asp Thr His 385 390 395 400Val Asn Lys Thr Tyr Pro Val Asp Met Ser Pro Thr Phe Thr Trp Lys 405 410415 Asn Ser Lys Leu Pro Asn Asp Ile Thr Gly Arg Gly Pro Gln Ile Leu 420425 430 Met Ile Ala Val Phe Thr Leu Thr Gly Ala Val Val Leu Leu Leu Leu435 440 445 Val Ala Leu Leu Met Leu Arg Lys Tyr Arg Lys Asp Tyr Glu LeuArg 450 455 460 Gln Lys Lys Trp Ser His Ile Pro Pro Glu Asn Ile Phe ProLeu Glu 465 470 475 480 Thr Asn Glu Thr Asn His Val Ser Leu Lys Ile AspAsp Asp Lys Arg 485 490 495 Arg Asp Thr Ile Gln Arg Leu Arg Gln Cys LysTyr Val Lys Lys Arg 500 505 510 Val Ile Leu Lys Asp Leu Lys His Asn AspGly Asn Phe Thr Glu Lys 515 520 525 Gln Lys Ile Glu Leu Asn Lys Leu LeuGln Ile Asp Tyr Tyr Thr Leu 530 535 540 Thr Lys Phe Tyr Gly Thr Val LysLeu Asp Thr Met Ile Phe Gly Val 545 550 555 560 Ile Glu Tyr Cys Glu ArgGly Ser Leu Arg Glu Val Leu Asn Asp Thr 565 570 575 Ile Ser Tyr Pro AspGly Thr Phe Met Asp Trp Glu Phe Lys Ile Ser 580 585 590 Val Leu Tyr AspIle Ala Lys Gly Met Ser Tyr Leu His Ser Ser Lys 595 600 605 Thr Glu ValHis Gly Arg Leu Lys Ser Thr Asn Cys Val Val Asp Ser 610 615 620 Arg MetVal Val Lys Ile Thr Asp Phe Gly Cys Asn Ser Ile Leu Pro 625 630 635 640Pro Lys Lys Asp Leu Trp Thr Ala Pro Glu His Leu Arg Gln Ala Asn 645 650655 Ile Ser Gln Lys Gly Asp Val Tyr Ser Tyr Gly Ile Ile Ala Gln Glu 660665 670 Ile Ile Leu Arg Lys Glu Thr Phe Tyr Thr Leu Ser Cys Arg Asp Arg675 680 685 Asn Glu Lys Ile Phe Arg Val Glu Asn Ser Asn Gly Met Lys ProPhe 690 695 700 Arg Pro Asp Leu Phe Leu Glu Thr Ala Glu Glu Lys Glu LeuGlu Val 705 710 715 720 Tyr Leu Leu Val Lys Asn Cys Trp Glu Glu Asp ProGlu Lys Arg Pro 725 730 735 Asp Phe Lys Lys Ile Glu Thr Thr Leu Ala LysIle Phe Gly Leu Phe 740 745 750 His Asp Gln Lys Asn Glu Ser Tyr Met AspThr Leu Ile Arg Arg Leu 755 760 765 Gln Leu Tyr Ser Arg Asn Leu Glu HisLeu Val Glu Glu Arg Thr Gln 770 775 780 Leu Tyr Lys Ala Glu Arg Asp ArgAla Asp Arg Leu Asn Phe Met Leu 785 790 795 800 Leu Pro Arg Leu Val ValLys Ser Leu Lys Glu Lys Gly Phe Val Glu 805 810 815 Pro Glu Leu Tyr GluGlu Val Thr Ile Tyr Phe Ser Asp Ile Val Gly 820 825 830 Phe Thr Thr IleCys Lys Tyr Ser Thr Pro Met Glu Val Val Asp Met 835 840 845 Leu Asn AspIle Tyr Lys Ser Phe Asp His Ile Val Asp His His Asp 850 855 860 Val TyrLys Val Glu Thr Ile Gly Asp Ala Tyr Met Val Ala Ser Gly 865 870 875 880Leu Pro Lys Arg Asn Gly Asn Arg His Ala Ile Asp Ile Ala Lys Met 885 890895 Ala Leu Glu Ile Leu Ser Phe Met Gly Thr Phe Glu Leu Glu His Leu 900905 910 Pro Gly Leu Pro Ile Trp Ile Arg Ile Gly Val His Ser Gly Pro Cys915 920 925 Ala Ala Gly Val Val Gly Ile Lys Met Pro Arg Tyr Cys Leu PheGly 930 935 940 Asp Thr Val Asn Thr Ala Ser Arg Met Glu Ser Thr Gly LeuPro Leu 945 950 955 960 Arg Ile His Val Ser Gly Ser Thr Ile Ala Ile LeuLys Arg Thr Glu 965 970 975 Cys Gln Phe Leu Tyr Glu Val Arg Gly Glu ThrTyr Leu Lys Gly Arg 980 985 990 Gly Asn Glu Thr Thr Tyr Trp Leu Thr GlyMet Lys Asp Gln Lys Phe 995 1000 1005 Asn Leu Pro Thr Pro Pro Thr ValGlu Asn Gln Gln Arg Leu Gln Ala 1010 1015 1020 Glu Phe Ser Asp Met IleAla Asn Ser Leu Gln Lys Arg Gln Ala Ala 1025 1030 1035 1040 Gly Ile ArgSer Gln Lys Pro Arg Arg Val Ala Ser Tyr Lys Lys Gly 1045 1050 1055 ThrLeu Glu Tyr Leu Gln Leu Asn Thr Thr Asp Lys Glu Ser Thr Tyr 1060 10651070 Phe

1. A vaccine composition comprising: a) a protein comprising at leastone epitope of human ST receptor protein or a nucleic acid molecule thatencodes said protein; and b) a pharmaceutically acceptable carrier ordiluent.
 2. The vaccine composition of claim 1 comprising said proteinwherein said protein an epitope of the extracellular domain of the humanST receptor protein.
 3. The vaccine composition of claim 2 comprisingsaid protein wherein said protein comprises the extracellular domain ofthe human ST receptor protein.
 4. The vaccine composition of claim 3comprising said protein wherein the protein comprises the human STreceptor protein.
 5. The vaccine composition of claim 4 comprising saidprotein wherein the protein consists of the human ST receptor protein.6. The vaccine composition of claim 1 comprising a nucleic acid moleculethat encodes said protein wherein said protein comprises an epitope ofthe extracellular domain of the human ST receptor protein.
 7. Thevaccine composition of claim 6 comprising a nucleic acid molecule thatencodes said protein wherein said protein comprises the extracellulardomain of the human ST receptor protein.
 8. The vaccine composition ofclaim 7 comprising a nucleic acid molecule that encodes said proteinwherein the protein comprises the human ST receptor protein.
 9. Thevaccine composition of claim 8 comprising a nucleic acid molecule thatencodes said protein wherein the protein consists of the human STreceptor protein.
 10. The vaccine composition of claim 1 comprising anucleic acid molecule that encodes said protein wherein said nucleicacid molecule is a plasmid.
 11. The vaccine composition of claim 1comprising a nucleic acid molecule that encodes said protein whereinsaid nucleic acid molecule is within a viral vector or a bacterial cell.12. The vaccine composition of claim 11 wherein said viral vector is arecombinant vaccinia virus.
 13. A haptenized protein comprising at leastone epitope of human ST receptor protein.
 14. The haptenized protein ofclaim 13 wherein said protein comprises at least one epitope of theextracellular domain of the human ST receptor protein.
 15. Thehaptenized protein of claim 14 wherein said protein comprises theextracellular domain of the human ST receptor protein.
 16. Thehaptenized protein of claim 15 wherein the protein comprises the humanST receptor protein.
 17. A vaccine composition that comprises ahaptenized protein of claim 13 and a pharmaceutically acceptable carrieror diluent.
 18. A vaccine composition comprising killed or inactivatedcells or particles that comprise a protein comprising at least oneepitope of human ST receptor protein and a pharmaceutically acceptablecarrier or diluent.
 19. The vaccine of claim 18 wherein said killed orinactivated cells or particles comprise a protein with an epitope of theextracellular domain of the human ST receptor protein.
 20. The vaccineof claim 19 wherein said killed or inactivated cells or particlescomprise a protein with the extracellular domain of the human STreceptor protein.
 21. The vaccine of claim 20 wherein said killed orinactivated cells or particles comprise the human ST receptor protein.22. The vaccine of claim 21 wherein said killed or inactivated cells orparticles comprise killed or inactivated colorectal tumor cells.
 23. Thevaccine of claim 18 wherein said killed or inactivated cells orparticles are haptenized killed or inactivated cells or particles.
 24. Amethod of treating an individual who has metastasized colorectal cancercomprising the step of administering to such an individual atherapeutically effective amount of a vaccine of claim
 1. 25. A methodof treating an individual who has been identified as being susceptibleto metastasized colorectal cancer comprising the step of administeringto such an individual a prophylactically effective amount of a vaccineof claim 1.